Product Highlights

Direct gel loading - eliminates the need for further processing following reaction completion

Product Description

BioMix™ Red is a complete ready-to-use 2x reaction mix containing a stable Taq DNA polymerase. It contains an additional inert red dye that permits easy visualization and direct loading onto a gel. There is no need to add loading buffer as the mix is of sufficiently high density to sink to the bottom of the gel. The red dye migrates like a 350 bp fragment on a 2% agarose TAE gel (or 600 bp on a 1% agarose).

BioMix Red has been developed to perform PCR assays of many common genomic and cDNA templates; the user has simply to add water, template and primers. It reduces the time required to set-up reactions, thereby minimizing the risk of contamination. Reproducibility is ensured by reducing the number of pipetting steps that can lead to errors.

BioMix Red is supplied with additional MgCl2 solution should any fine adjustments be required.

Specification

Components

Volume

BIO-25005: 100 x 50 µL Reactions: 2 x 1.25 mL

BIO-25006: 500 x 50 µL Reactions: 10 x 1.25 mL

Concentration

2x

Storage & Stability

All components should be stored at -20°C upon receipt for optimum stability. Repeated freeze/thaw cycles should be avoided. When stored under the recommended conditions and handled correctly, full activity of the reagents is retained until the expiry date indicated on the outer box label.

Alternatively, BioMix Red can be stored for up to up to 2 weeks at +4°C.

Shipping conditions

On Dry Ice or Blue Ice.

FAQs

No, the dyes and composition of the Red Reaction Buffers and Mixes are such that the samples will sink easily into the well and the samples can be clearly seen, thus no loading buffer is required to load your samples on an agarose gel.

The dye present in the Red Mixes or Red Reaction Buffer does not interfere with PCR product isolation procedures or subsequent applications like sequencing. An exception is the quantification of PCR products in colored buffers with photometric methods or fluorescence assays. We cannot exclude the impact of the dye on quantification results and suggest the purification of these samples, for instance with ISOLATE II PCR & Gel kit or SureClean Plus.

Bioline's mixes contain magnesium chloride at the following concentrations (please see table below). An additional tube of 50 mM MgCl2 solution is supplied with each mix to facilitate further optimization if required.

PCR can be a challenging technique, with various parameters to optimize to achieve the best results. If you are having problems, these could be easily resolved by addressing a few issues. Please see our PCR troubleshooting guide for suggestions and help with your specific problems.

These terms refer to parameters to be considered when performing PCR and are important features in selecting the correct enzyme for your needs. Understanding what they mean is therefore crucial:
Yield: The amount of DNA produced in a PCR reaction.
Fidelity: The accuracy of the enzyme at incorporating the correct dNTP to the elongating DNA strand.
Processivity: The length of time a polymerase is associated with the template and therefore the size of fragment which can be amplified.
Specificity: A measure of the unwanted by-products generated in a reaction.

All Bioline polymerases are available in the convenient format of a 2x master mix, containing all the reagents and additives necessary to perform successful PCR, with the exception of template, primers and water.
This formulation not only provides a convenient and hassle-free PCR setup, but also significantly reduces the chances of human error, inaccuracies and contamination by reducing the pipetting steps required.

Whilst the exact composition of the mixes is proprietary information, all mixes are made up of reaction buffer, magnesium, dNTPs, polymerase and additives, designed to keep the polymerase stable in the mix, as well as provide the optimal conditions for PCR.

The MyTaq HS DNA polymerase works very well with difficult samples like GC rich or bisulfite converted DNA. Especially the hot-start is important, as it decreases primer dimer and unspecific amplification. We would suggest to start with the recommended protocol. If optimization is needed, we would suggest to optimize the annealing temperature. Due to the degraded DNA after bisulfite conversion it may be useful to increase the amount of template and polymerase.

If possible, we would recommend an additional cleanup of affected samples, for instance, customers had very good experiences with the clean-up of samples containing humic acids with SureClean Plus. Alternatively decreasing the template concentration will help to minimize the amount of inhibitors. The amount of primer can be increased, but do not exceed the suggested limits.